INTRODUCTION:

Microalgae have been widely used as nutraceuticals (drugs and supplements) various types of microalgae have been studied to have bioactive compounds that are useful as drugs and supplements including; Scenedesmus dimorphus,

Parachlorella kessleri, Spirulina platensis, chorella vulgaris, Dunaliella, Chroococcus, Nanochloropsis and others. S. platensis (S.platensis) is included in microalgae which contains high protein, namely 55-70%, good fats (omega-3 and omega-6 and PUFA), carbohydrates, biopigments (Phycocyanin, astaxanthin, beta-carotene, luetein, chlorophyll and carotenoids)¹². As a future food because it has suitable nutritional content as functional food, because the amount of protein contained in S. platensis is quite high and the amino acids that are often found in S. platensis protein are essential amino acids such as methionine (1,3-2,75 %), cysteine (0.5-0.75%), tryptophan (1-1.95%), and lysine (2.6-4.63%). The amount of poly unsaturated fatty acids (PUFAs) in S. platensis is about 1.3-15% of the total fat, which is 6-6.5%. The highest fatty acids are gamma linoleic acid (GLA) which is about 25-60% of the total fat, then palmic acid (44.6-54.1%), oleic acid (1-15.5%), and linoleic acid (10.8-30.7%)³. The content of vitamins contained in S. platensis such as vitamin A, vitamin B(B1, B2, B3, B6, B9, B12), vitamin C, vitamin D, and vitamin E.⁴

Spirulina sp. used as food health and therapeutic has been commercialization in several countries due to its constituents, especially protein, vitamins, and a rich source of pigments such as Phycocyanin⁵. Based on the nature of polarity, the polar pigments contained in S. platensis amounted to about 42.272±0.05mg/g of the total dry weight, where this polar pigment is a phycobiliprotein or phycobilin group consisting of allo Phycocyanin (APC, red); phycoerythrin (PE, pink/purple); phycoerythrocyanin (PEC, orange); and Phycocyanin (C-PC, blue-green), which contains different chromophore groups such as phycoerythrobilin (PEB, red); phycobiliviolin (PXB, purple); phycourobilin (PUB, yellow); and phycocyanobilin (PCB, purple)⁶. Meanwhile, the non-polar pigment content was 4.498±0.06mg/g of the total dry weight consisting of carotenoids (carotene and xanthophyll) and chlorophyll a

Phycocyanin is the main pigment found in S. platensis where the amount is around 6.7-11.7%⁷. Phycocyanin belongs to the group of phycobiliprotein polar pigments that give a blue-green color because it contains an open-chain tetrapyrrole chromophore group (phycocyanobilin) which is covalently bound to the Apo protein molecule⁸. Phycocyanin is a pigment that has very low stability, its colour will fade at temperatures above 45℃ and pH below 4 (the optimum pH of Phycocyanin is 4-9). Phycocyanin pigments are able to capture radiation from sunlight so they are very susceptible to environmental conditions such as heat, light, and oxygen. Phycocyanin functions as antiplatelet, hepatoprotective, antioxidant, anti-inflammatory, reducing cholesterol levels, anticancer, and antidiabetic⁴. Oxidation is a chemical reaction that transfer electrons from a substance to an oxidizing agent⁹. Interestingly, Phycocyanin has open-chain tetra pyrrole chromophore molecules that can be used as an antioxidant. The mechanism of action is by donating a hydrogen atom bonded to C-10 in a tetra pyrrole molecule. The mechanism of Phycocyanin against oxidative stress in the body is by increasing levels of GSH, CAT, and glucose-6-phosphate dehydrogenase (G6PD). Phycocyanin is able to prevent a decrease in enzymatic antioxidant levels in the body exposed to oxidative stress. In addition, Phycocyanin can also inhibit the occurrence of lipid peroxidation due to free radicals and restore antioxidant function that has decreased due to free radicals. Peroxidation of lipids can greatly alter the physicochemical propertied of membrane lipid bilayer resulting in severe cellular disfunction¹⁰^,¹¹. The ability of Phycocyanin to inhibit lipid peroxidation is by binding to hydroxyl radicals so that it can suppress lipid peroxidation¹². The main applications of phycobiliproteins are as fluorescent markers of cells and macromolecules in biomedical research and the most sensitive fluorescence techniques due to the high molar absorptivity of C-PC and other phycobiliproteins at visible light wavelengths¹³.

Phycocyanin has significant antioxidant, anti-inflammatory, cardio protective, and radical scavenging properties even used in food colouring and cosmetics because it is non-toxic and non-carcinogenic¹⁴. Inflammation is a complex biological response of body tissues to noxious stimuli characterized by an increase in the ability of cells to produce pro-inflammatory cytokines such as interleukin-6 (IL-6), interleukin-1 (IL-1) and tumor necrosis factor-α (TNF-α)¹⁵^,¹⁶ Pro-inflammatory interacts with genetic background and environmental factors. Anti-inflammatory compounds can inhibit the ROS-induced activation of the transcription factor nuclear factor-κB (NF-B), which then effectively suppresses the production of inflammatory cytokines such as IL-1β, IL-6, and TNF-α. TNF is a major mediator of inflammation, which +can stimulate fibroblast growth. TNF can destroy blood vessels but also causes angiogenic factors¹⁷ . The present work describes was to conducted isolation and identification of microalgae Spirulina platensis isolated from Lake Maninjau, West Sumatra, Indonesia. Additionally, we investigate characteristics and the activity of Phycocyanin extracted from Spirulina platensis as antioxidant and anti-inflammatory.

MATERIALS AND METHODS:

Materials:

Spirulina platensis, extract of Phycocyanin, monosodium glutamate (MSG) (Merck), Zarrouk Growth media-Spirulina nutrition, ABTS reagent, phosphate buffer saline (PBS), Trolox, Tris HCl buffer pH 7.4, DMSO, linoleic acid, lipoxygenase enzyme (type V) (Thermo Fisher Scientific), FOX reagen: sulfuric acid (30mM), xylenol orange (100M), iron (II) sulphate (100M), and methanol/water (9:1) (Sigma Chemicals).

Instrument:

Centrifugation (H-C-12), vortex (VELP®scientifica Zx3), Freeze-dryer (Buchi, Lyovapor L-200), Autoclave, UV-Vis Spectrophotometer (Genesys 1280 Serial No A120657), FTIR, and Plate ELISA reader.

Microalgae Isolation and Idenitification:

Freshwater samples were collected from Maninjau Lake, West Sumatra, Indonesia. Water samples filtered by the plankton net were placed into a bottle filled with sterile Zarrouk’s medium (ZM) for further identification and isolation. The samples were inoculated into 1000ml of in Erlen Mayer flasks. The samples were incubated in the growth chamber having a facility to maintain a light intensity of 3000-3200 lux with at room temperature (25-28°C). Serial dilution combined with micropipette washing technique method was used to isolate microalgae in this study². The isolated microalgae identified as Spirulina Platensis. Individual microalgae colonies were identified using a microscope. The microalgae samples were submitted for the DNA extraction and purification process using the Dneasy ® Tissue Kit (Qiagen Sciences, Valencia, Calif., Md, USA). After DNA isolation and purification, the 18S rDNA genomic region amplification was carried out using pairs of universal primers. Then, 18s rRNA was used for DNA sequencing and compared with the GenBank database using BLAST. The sequences were submitted to distance estimation, and the phylogenetic tree was built according to Neighbor- Joining (NJ) method using MEGA11¹⁸.

Phycocyanin Cultivation and Production:

S. platensis was cultured in Zarrouk's medium, treated by adding 7.5mM monosodium glutamate (MSG) to the medium as metabolic stress to increase Phycocyanin production in S. platensis culture. The addition of MSG was carried out at the beginning of cultivation. Cultures were sampled every 2 days until the final stationary phase and each sample was measured by optical density (OD₆₈₀) to determine the growth curve and optimum concentration of the Phycocyanin.

(OD₆₂₀ – 0.474 × OD₆₅₂

Concentration of Phycoxyanin = --------------------------------

5.34

Antioxidant Activity:

The antioxidant activities assay ABTS methods. Pipe 1 mL of ABTS stock solution, then the volume was made up to 25mL with PBS solution pH 7.4 in a volumetric flask. Then the absorbance was measured at a wavelength of 734nm (absorption value 0.7±0.02). Sample Solution Preparation: The dry extract of Phycocyanin was weighed at 25mg and then dissolved using deionized water in a 25mL volumetric flask. Then it was diluted at concentrations, 20, 40, 60, 80, and 100 mg/L respectively. Each solution was pipetted 0.1mL and added 2mL of ABTS stock solution. They incubated for 6 minutes in a dark room. The preparation of the stan Trolox stock solution with a concentration of 100mg/L in 25mL. Then diluted at concentration was varied, namely, 2, 4, 6, 8, and 10mg/L. Each solution was then pipetted 0.1mL and put into a vial. Then, to each vial, 2 mL of ABTS solution was added and homogenized. The absorbance of control solution, sample, and positive control was measured using a microplate reader at a wavelength of 734nm. Then the value of the inhibition is calculated using the equation:

A control – A sample

% Inhibition = ------------------------------------------- × 100

A Control

Anti-inflammation Test:

Activity of Lipoxygenase:

FOX test based on a 96-well micro-plate for measurement of lipoxygenase. An aliquot of 50µL LOX was dissolved in 50mM Tris HCl buffer, pH 7.4 so that the final concentration, 100ng protein/mL, and then added 20µL of the test sample (Phycocyanin or standard inhibitor) in each well of the 96-well micro-plate and incubated at 25°C for 5 minutes. Solution of LOX as control ware pipette 50µL and 20µL of buffer containing 0.2% v/v DMSO into the well. Blank containing LOX enzyme and incubation, after, that add FOX reagent, and then linoleic acid as substrate. The initiation reaction was by the addition of 50µL of linoleic acid (final concentration, 140M) in 50mM. Tris HCl buffer, pH 7.4, and the reaction mixture were incubated at 25°C for 20 min in the dark. Final concentrations of reaction mixture of LOX and linoleic acid on a total volume of 120µL for the reaction mixture. The assay ended with the addition of 100µL of freshly prepared FOX reagent. After termination, the Fe³⁺–dye complex is allowed to develop for 30 min at 25°C before being measured at 560nm on a micro-plate reader

(A_Control – A_Blank) – (A_Sample - A_Blank)

Concentration of Lipoxygenase = ------------------ ×100%

(A_Control – A_Blank)

Inhibition of BSA denaturation:

The anti-inflammatory activity test uses the Mizushima and Kobayashi methods. Bovine serum albumin adjusts at pH 6.3 by adding a small amount of hydrochloric acid. An aliquot of serum 0.45mL adds to 0.05mL of Phycocyanin. The mixture was incubated at room temperature for 20 minutes, then heated at 57^oC for 3 minutes to denature the protein. After the mixture has cooled, add 2.5mL of saline solution with phosphate buffer (0.07 M Na3PO4 0.15 M NaCl) pH 6.3. The solution measurement used spectrophotometer at 660 nm.

Phycocyanin Characterization:

UV-Vis Spectrophotometer:

Identification of phycobiliprotein pigments from S. platensis extract was used by measuring the phycobiliprotein spectrum at a wavelength of 300-700 nm. The maximum absorption of the protein bound to Phycocyanin, and the type of Phycocyanin pigment present in samples are visible from the peak on the spectrum.

FTIR (Fourier Transform Infrared):

Identification and characterization using IR spectroscopy to determine the main functional groups in the Phycocyanin S. platensis. A sample of 20 mg was ground homogeneously with 10mg of potassium bromide. The mixture pressed with a strength of 10 tons/cm to form a thin and transparent pellet powder absorbance was measured¹⁹.

RESULT:

Microalgal identification:

Morphological examination of the isolated microalgae showed It has a bluish color (blue green algae) with an elongated spiral shape celled colony (Figure 1). Phylogenetic tree was constructed based on the BLAST from NCBI then analysis and phylogenetic tree that produced using MEGA11 software and based on Neighbor-Joining (NJ) method. gave the result that strain Microalgae AU-2 has highest similarity with other species namely Arthospira platensis PCC9108, Arthospira platensis A, and Spirulina platensis with sequence identity of all was 100% (Figure 2).

Figure 1: The results of studying the morphological features of the isolated microalgae (Spirulina platensis)

Figure 2: Neighbor-joining (NJ) phylogenetic tree for Microalgae AU-2 based on using 18S rRNA sequences

FTIR Spectra of the Microalgae Biomass:

The Fourier Transform Infrared (FTIR) spectroscopy provides information on the chemical structure of microalgae biomass by identifying the peaks of its spectra, which can indicate the presence of functional groups. FTIR spectroscopy provides information about the chemical structure of Spirulina platensis biomass by identifying its spectral peaks, which are produced by functional group vibrations. The FTIR spectra of Spirulina platensis biomass are showed in Figure 3.

Figure 3: The results of studying FTIR spectra of microalgae biomass Spirulina platensis AU-2

Phycocyanin Production:

S. platensis microalgae were isolated from Maninjau Lake and identified by molecular biology. Obtained isolate local S. platensis from Maninjau Lake, which has potential as a Phycocyanin producer. Increased production of Phycocyanin from S. platensis was induced with stress metabolites using monosodium glutamate. Administration of monosodium glutamate in S. platensis growth media as metabolic stress can increase Phycocyanin production. MSG with a concentration of 7.5mM increased Phycocyanin production twofold compared to no MSG. Phycocyanin production was obtained from S. platensis with MSC 7.5 mM (48.7±0.443mg/L) and without MSG (29.68±0.364 mg/L) at an incubation time of 16 days (Figure 4).

Figure 4: Phycocyanin production with 7.5 mM MSG as metabolic stress and without MSG

Phycocyanin Characterization by UV-Vis Spectrophotometer:

Phycocyanin belongs to phycobiliprotein polar pigments that give a green-blue colour because it contains an open-chain tetra pyrrole chromophore group (Phycocyanobilin) covalently bound to the apoprotein molecule. Phycocyanin are pigment-protein complexes of the phycobiliprotein family. Phycobiliproteins consist of two subunits (alpha and beta) with a protein backbone covalently bound to 1-2 linear tetra pyrrole chromophores. Spirulina platensis consisted of two types of Phycocyanin namely, C-Phycocyanin and A-Phycocyanin bound to this protein shown from the character of UV-Vis. Characterization of Phycocyanin with UV-Vis showed maximum wave absorption of three peaks, namely C-Phycocyanin is 615 nm, A-Phycocyanin, shows the maximum absorption of 652 nm and 280 nm indicating the total protein uptake bound to Phycocyanin, as shown in Figure 5.

Figure 5: Spectrum and maximum wavelength of Phycocyanin extract from Spirulina platensis (C-Phycocyanin 620 nm, A-Phycocyanin 652 nm, and Protein bound to Phycocyanin 280 nm)

FTIR (Fourier Transform Infrared) of Phycocyanin:

Analysis by FTIR, the sample is exposed to infrared (IR) radiation. IR radiation impacts the atomic vibrations of a molecule in the sample, resulting in the absorption and transmission of certain energies. FTIR is used to determine the specific molecular vibrations contained in the samples. FTIR spectrum of the Phycocyanin extract from Spirulina platensis shown in Figure 6 and Table 1 shows the functional groups with frequency quantified.

Figure 6: The FTIR spectrum of Phycocyanin extracted from Spirulina platensis with MSG as metabolic stress was compared with the standard

Table 1: The peak in the functional group from Phycocyanin with MSG in the FTIR spectrum compared to standard

Functional Groups

Reference Wavenumber

Wavenumber

Standard Phycocyanin

Phycocyanin with MSG

Secondary Amide

(CO-N-R)

N-H-R

O-H

(3500-3310)

C=O

(1680-1620)

3312.18

1620.56

3334.44

1642.13

Secondary Amide

R-N-H

C-N

(1220-1020)

1065.94

1079.94

Pyrrol ring

(1600-1300)

1451.77

1465.73

Antioxidant Test with ABTS Method:

Determination of antioxidant activity based on the ABTS method in principle is the removal of the color of the ABTS cation radical from turquoise to colourless. The percentage value of ABTS•+ radicals reduction by Phycocyanin extract from S. platensis shown in Figure 7

Figure 7: Phycocyanin inhibition of ABTS free radicals compared with Trolox as standard

Phycocyanin Inflammatory Activity by BSA Denaturation Method:

Phycocyanin activity as an anti-inflammatory with BSA protein denaturation inhibition method obtained IC50 of 62.3 ppm compared to standard Aspirin IC50 of 42.17 ppm. Inhibition of Phycocyanin on BSA denaturation at a concentration of 62.3 mg/mL effectively inhibited 50% of BSA protein denaturation (Figure 8).

Figure 8: Inhibitory Activity of Inflammation by Phycocyanin compared to Aspirin

Lipoxygenase Enzyme Activity:

Phycocyanin activity as anti-inflammatory based on inhibition the lipoxygenase enzyme activity test has IC₅₀ of 48.73ppm, with a small value of 50ppm showing that Phycocyanin is a strong inhibitor of the lipoxygenase (LOX) enzyme. The results of the inhibition of lipoxygenase enzymes with various concentrations of Phycocyanin concentrations are shown in the figure 9.

Figure 9: Inhibition of lipoxygenase enzymes with various concentrations of 20 – 120 ppm

DISCUSSION:

This was identified as Spirulina platensis and have morphological characters of these cultures are similar to that of the reference strain microalgae AU-2 with less spiraling and blunt ends with florescence green appearance²⁰. The Microalgae AU-2 strains present morphological characteristics that suggest that they probably belong to the genus Cyanobacteria. Microalgae AU-2 strain was characterized using 18 rRNA gene analysis and its phylogenetic relationship with others of cyanobacteria species in NCBI. Phylogenetic tree was constructed based on the BLAST from NCBI then analysis and phylogenetic tree that produced using MEGA11 software and based on Neighbor-Joining(NJ) method gave the result that strain Microalgae AU-2 has highest similarity with other species namely Arthospira platensis PCC9108, Arthospira platensis A, and Spirulina platensis with sequence identity of all was 100% (Figure 2). By molecular analysis of the amplified sequences of the 18SrDNA of the Microalgae AU-2 strains by BLAST/NCBI, high similarity with species of the genus Cyanobacteria such as Arthospira platensis or Spirulina platensis was observed.

The presence of functional groups in the FTIR spectrum, in Figure 6 shows that microalgae biomass contains a number of functional groups that differ from one and another species of microalgae. The FTIR spectrum, which indicates the presence of functional groups will be identified and associated with the presence of macromolecules such as lipids, proteins and carbohydrates over the wave number of range 4000-500 cm^-1²¹. Infrared spectrum analysis results of Spirulina platensis microalgae biomass showed seven distinct bands at 3440, 1645, 1384 and 1084 cm^-1, which revealed microalgae biomass had or contains functional organic groups. Lipids are characterized by a strong vibrational band at 1599-1647 cm^-1 (C = O stretch) and a weaker band at 1462 cm^-1(CH₂), while proteins are characterized by two strong and broad bands at 1640 cm^-1 (amide I: C = O stretch) and 1535 cm^-1 (amide II: NH deformation and C–N stretch). Carbohydrates have strong absorption in the 1200–900 cm^-1 region (C–O–C and C–OH stretch) including some characteristic bands for certain types of carbohydrates, such as cellulose (at 1107, 1055 and 1028 cm^-1) and amylose (at 1076 cm^-1) ^22,23. Meanwhile, the spectrum at 3424-3440 cm^–1 reveals the presence of hydroxyl group. Microalgae Spirulina platensis contains protein and carbohydrate content which was indicated due to the presence of high carbohydrates and proteins spectrum in 1384 cm^-1 ^24,25. FTIR band spectra as a whole confirm the presence of three main constituents in biomass, namely carbohydrates, proteins, and lipids. These three components are macromolecular compositions that play an important role in the development of microalgae as a source of food and health.

Antioxidant activity Phycocyanin was done by BTS method. Trolox was used as a comparison or standard solution because it was soluble in water. The IC₅₀value of Trolox is 18.77ppm, classified into a powerful antioxidant. Percent Phycocyanin inhibition against ABTS free radicals was determined based on the concentration of Phycocyanin which could inhibit free radicals by 50% (IC₅₀). Phycocyanin has an IC₅₀ = 46.32 ppm, small than 50ppm, and includes a powerful antioxidant.⁵^,²⁶^,²⁷^.Phycocyanin activity as an anti-inflammatory with BSA protein denaturation inhibition method obtained IC₅₀ of 62.3ppm compared to standard Aspirin IC₅₀ of 42.17ppm. This study was larger than the results of research on the red algae extract, G. folifera, namely the average inhibition was 58.7ppm²⁸. Nevertheless, the value was smaller than the results of research on seeds extract of Morinda citrifolia and Malvastrum coromandelianum leaf extracts, namely the average inhibition were 72.69 and 89.17 respectively²⁹^,³⁰. Another previous study revealed that appreciable antioxidant activity of phycocyanin. At a concentration of 200μg/mL, the phycocyanin exhibited a maximum absorbance of 0.49 by phosphomolybdenum assay, 0.85 absorbance by ferric ion reducing assay. The results clearly indicate that the phycocyanin can act as an electon donor and reduce the oxidized intermediates of lipid peroxidation processes. From these results we can conclude that phycocyanain can be a promising primary and secondary antioxidant compound³¹. Compounds that inhibit protein denaturation by more than 20% are also considered to have anti-inflammatory activity and can be a reference value for drug development. Then, previous study reported extract water Spirulina platensis have potential as antiperoxidative³²^,³³. These results indicate that the Phycocyanin extract has anti-inflammatory activity. Compounds that can stabilize proteins during the protein denaturation process are compounds that have the potential to an anti-inflammatory. The interaction between Phycocyanin and BSA where formed bonds, Phycocyanin whit tyrosine, threonine, and lysine from BSA. The bond between Phycocyanin and anima acid from BSA (tyrosine, threonine, and lysine) stabilizes the protein and prevents BSA denaturation¹⁴^,³⁴.

Based on the results obtained, inhibition of the lipoxygenase enzyme to determine Phycocyanin as an anti-inflammatory has IC₅₀ of 48.73 ppm, with a small value of 50 ppm showing that Phycocyanin is a strong inhibitor of the lipoxygenase (LOX) enzyme. This study is also in line with testing the anti-inflammatory activity carried out by induction of carrageenan and cotton pellets granuloma model on ethanoic fraction of Thuja occidentalis which showed significant results³⁵. The ability of Phycocyanin to inhibit LOX activity stated Phycocyanin is beneficial as an anti-inflammatory. Additionally, the other study explains that the PC-LOX interaction is of a function-freezing, protein-protein interaction in nature. The wide spectrum of properties of PC might be due to its function as a powerful protein hub showing non-specific protein-protein interactions³⁶. Lipoxygenases are present in the human body and play a role in stimulating inflammatory reactions. Excessive amounts of reactive oxygen species can cause inflammation that stimulates the release of cytokines and subsequent activation of LOX. Inflammation links many diseases, such as cancer, stroke, and cardiovascular and neurodegenerative diseases³⁷.

The test used in this study measures the conversion of linoleic acid to linoleic hydro peroxide in the presence of LOX. Linoleic hydro peroxide then oxidizes Fe²⁺ in FOX reagent to Fe³⁺ ions, which then interact with acidified xylene orange to produce Fe³⁺ a complex dye that absorbs light at 560 nm. Phycocyanin extract as a LOX inhibitor will reduce the formation of the Fe³⁺dye complex.³⁷^,³⁸ Phycocyanin has primarily dietary nutrition and supplements and is a potent anti-inflammatory, anticancer, antiviral and neuroprotective, and hepatoprotective.³⁹^,⁴⁰ Oxidative stress is a primer source of inflammation, leading to the loss of dopaminergic neurons resulting in neurological disorders such as Alzheimer's disease, cancer, aging and variety of diseases⁴¹^,⁴²^,⁴³. The micro molar concentration of Phycocyanin reduced the concentration of proxy radicals by half, indicating its activity as antioxidant diseases.

CONCLUSION:

Spirulina platensis isolated from the waters of Lake Maninjau has potential as a producer of Phycocyanin. Administration of MSG as a stress metabolite can increase production twofold, at (48.7±0.443mg/L) to (29.68±0.364 mg/L) for the control. Characterization with UV-Vis showed that the absorption at a wavelength of 620 nm was Phycocyanin. Antioxidant activity of Phycocyanin obtained IC₅₀ is 46.32ppm, using standard Trolox where IC₅₀ of 18.773ppm, while the activity of Phycocyanin as anti-inflammatory with the BSA protein denaturation inhibition method observed an IC₅₀ of 62.3 ppm compared to the standard Aspirin IC₅₀ of 42.17ppm, The Inhibition of lipoxygenase enzyme obtained IC₅₀ of 48.73ppm. Characterization of the extract from S. platensis by UV-Vis and FTIR indicated Phycocyanin. Therefore, it can be concluded that Phycocyanin is a good agent as an antioxidant and anti-inflammatory.

CONFLICT OF INTEREST:

The authors have no conflicts of interest regarding this investigation.

ACKNOWLEDGMENTS:

The Authors thanks the Ministry of Research, Technology and Higher Education of the Republic of Indonesia and Andalas University, Padang Indonesia for fund this research through Percepatan Guru Besar (PGB) scheme.

REFERENCES:

1. Armaini, Salim M, Pribadi P. Induction effect of microalgae Scenedesmus dimorphus against hematology on mice (Mus musculus) suffering anemia diseases. Asian J. Pharm. Clin. Res. 2018; 11(7): 348-352. doi:10.22159/ajpcr.2018.v11i7.24825

2. Kusnanda AJ, Dharma A, Armaini, Syafrizayanti, Chaidir Z, Pardi H. Antioxidant and α-glucosidase inhibitor activity characterization of bioactive components from Parachlorella kessleri AUP5. AACL Bioflux. 2023; 16(4): 2252-2263.

3. Rinaldi R, Armaini, Salim M. A Selection of Nitroen Source for Biomass and Lipid Production of Snecedesmus dimorphus Microalgae. Researcj J. Pharm Biol Sci. 2(3): 847-855.

4. Koru E. 7. Earth Food Spirulina (Arthrospira).pdf.

5. Wu HL, Wang GH, Xiang WZ, Li T, He H. Stability and Antioxidant Activity of Food-Grade Phycocyanin Isolated from Spirulina platensis. Int J Food Prop. 2016; 19(10): 2349-2362. doi:10.1080/10942912.2015.1038564

6. Munier M, Morançais M, Dumay J, Jaouen P, Fleurence J. One-step purification of R-phycoerythrin from the red edible seaweed Grateloupia turuturu. J Chromatogr B Anal Technol Biomed Life Sci. 2015; 992: 23-29. doi:10.1016/j.jchromb.2015.04.012

7. Li Y, Zhang Z, Paciulli M, Abbaspourrad A. Extraction of phycocyanin—A natural blue colorant from dried spirulina biomass: Influence of processing parameters and extraction techniques. J Food Sci. 2020; 85(3): 727-735. doi:10.1111/1750-3841.14842

8. Chew KW, Chia SR, Krishnamoorthy R, Tao Y, Chu DT, Show PL. Liquid biphasic flotation for the purification of C-phycocyanin from Spirulina platensis microalga. Bioresour Technol. 2019; 288(May). doi:10.1016/j.biortech.2019.121519

9. Jangdey MS, Gupta A, Sah AK, Daharwal SJ. Role of antioxidants in developing novel delivery systems as longevity therapy. 2014; 6(3): 119-127.

10. Madhikarmi NL, Siddalinga KR. Study of oxidative stress and antioxidants status in iron deficient anemic patients. 2012; 4(4): 162-167.

11. Gunalan G, Suresh Kumar M, Sangeetha N. Preliminary phytochemical analysis and in vitro oxidant scavenging activity of Rosemary officinalis. Res. J. Pharm. Technol. 2011; 4(10): 1588-1590.

12. Grover P, Bhatnagar A, Kumari N, Narayan Bhatt A, Kumar Nishad D, Purkayastha J. C-Phycocyanin-a novel protein from Spirulina platensis- In vivo toxicity, antioxidant and immunomodulatory studies. Saudi J Biol Sci. 2021; 28(3): 1853-1859. doi:10.1016/j.sjbs.2020.12.037

13. Kuddus M, Singh P, Thomas G, Al-hazimi A. Recent Developments in Production and Biotechnological Applications of C-Phycocyanin. 2013; 2013.

14. Romay C, Gonzalez R, Ledon N, Remirez D, Rimbau V. C-Phycocyanin: A Biliprotein with Antioxidant, Anti-Inflammatory and Neuroprotective Effects. Curr Protein Pept Sci. 2005; 4(3): 207-216. doi:10.2174/1389203033487216

15. Landskron G, De La Fuente M, Thuwajit P, Thuwajit C, Hermoso MA. Chronic inflammation and cytokines in the tumor microenvironment. J. Immunol Res. 2014; 2014. doi:10.1155/2014/149185

16. Franceschi C. Inflammaging as a Major Characteristic of Old People: Can It Be Prevented or Cured? Nutr Rev. 2007;65(SUPPL.3):12-15. doi:10.1111/j.1753-4887.2007.tb00358.x

17. Balkwill FR, Mantovani A. Cancer-related inflammation: Common themes and therapeutic opportunities. Semin Cancer Biol. 2012; 22(1): 33-40. doi:10.1016/j.semcancer.2011.12.005

18. Kusnanda AJ, Dharma A, Chaidir Z. Carotenoid Profile of Freshwater Microalgae Mychonastes racemosus AUP1 and its Antioxidant properties. 2023; 16(January):1-7.

19. Musifa E, Kusnanda AJ, Dharma A, Sciences N, Andalas U, Limau K. Monosodium Glutamate (MSG) as Metabolic Stressors Stimulate the Production of Valuable Compounds in Spirulina platensis. 2023; 27(2): 731-743.

20. Singh NK, Dhar DW. Phylogenetic relatedness among Spirulina and related cyanobacterial genera. World J Microbiol Biotechnol. 2011; 27(4): 941-951. doi:10.1007/s11274-010-0537-x

21. Gopinatha S, Nagarajanb N. Journal of Applied Research and Technology. J Appl Res Technol. 2015; 13: 374-381.

22. Batcioʇlu M, Zimmermann B, Kohler A. A multiscale vibrational spectroscopic approach for identification and biochemical characterization of pollen. PLoS One. 2015; 10(9). doi:10.1371/journal.pone.0137899

23. Manjunatha SS, Girisha ST. Characterization of microalgal biomass through fourier transforms infrared (FT-IR) spectroscopy. Int J Bot Stud. 2021; 6(1): 57-60. https://www.researchgate.net/publication/348900141_Characterization_of_microalgal_biomass_through_fourier_transforms_infrared_FT-IR_spectroscopy

24. Assunção MFG, Amaral R, Martins CB, et al. Screening microalgae as potential sources of antioxidants. J. Appl Phycol. 2017; 29(2): 865-877. doi:10.1007/s10811-016-0980-7

25. Wang L, Wang L, Manzi HP, et al. Isolation and screening of Tetradesmus dimorphus and Desmodesmus asymmetricus from natural habitats in Northwestern China for clean fuel production and N, P removal. Biomass Convers Biorefinery. Published online 2020. doi:10.1007/s13399-020-01034-z

26. Safari R, Amiri RZ, Esmaeilzadeh Kenari R. Antioxidant and antibacterial activities of C-phycocyanin from common name Spirulina platensis. Iran J Fish Sci. 2020; 19(4): 1911-1927. doi:10.22092/ijfs.2019.118129

27. Qiao BW, Liu XT, Wang CX, Song S, Ai CQ, Fu YH. Preparation, Characterization, and Antioxidant Properties of Phycocyanin Complexes Based on Sodium Alginate and Lysozyme. Front Nutr. 2022; 9(May): 1-11. doi:10.3389/fnut.2022.890942

28. Jenifer P, Balakrishnan CP, Pillai SC. In-vitro Antioxidant activity of Marine Red Algae Gracilaria foliifera . Asian J. Pharm. Technol. 2017; 7(2): 105. doi:10.5958/2231-5713.2017.00018.6

29. Pal R, Girhepunje K, Shrivastava N, Hussain MM, Thirumoorthy N. Antioxidant and free radical scavenging activity of ethanolic extract of Morinda citrifolia. Res. J. Pharm. Technol. 2011; 4(8): 1224-1226.

30. Yadav AR, Mohite SK. Antioxidant activity of Malvastrum coromandelianum leaf extracts. Res J Top Cosmet Sci. 2020; 11(2): 59-61. doi:10.5958/2321-5844.2020.00010.2

31. Renugadevi K, Valli Nachiyar C, Sowmiya P, Sunkar S. Antioxidant activity of phycocyanin pigment extracted from marine filamentous cyanobacteria Geitlerinema sp TRV57. Biocatal Agric Biotechnol. 2018; 16(March): 237-242. doi:10.1016/j.bcab.2018.08.009

32. Chew KW, Yap JY, Show PL, et al. Microalgae biorefinery: high value products perspectives. Bioresour Technol. Published online 2017. doi:10.1016/j.biortech.2017.01.006

33. Ray S. Exploring the protective role of water extract of Spirulina platensis on flutamide-induced lipid peroxidation using 4-hydroxy nonenal and nitric oxide as model markers. Res. J. Pharm. Technol. 2011; 4(12): 1857-1860.

34. Liu R, Qin S, Li W. Phycocyanin: Anti-inflammatory effect and mechanism. Biomed Pharmacother. 2022; 153(July): 113362. doi:10.1016/j.biopha.2022.113362

35. S.K.Dubey, Batra A. Study of anti oxidant and anti-inflammatory activity from ethanol fraction of Thuja occidentalis Linn. Res. J. Sci. Technol. 2009; 1(1): 39-42.

36. Prasanth S, Kumar Arun G, Haridas M, Sabu A. Phycocyanin of marine Oscillatoria sp. inhibits lipoxygenase by protein-protein interaction-induced change of active site entry apace: A model for non-specific biofunctions of phycocyanins. Int. J. Biol Macromol. 2020; 165: 1111-1118. doi:10.1016/j.ijbiomac.2020.09.238

37. Jensen GS, Attridge VL, Beaman JL, Guthrie J, Ehmann A, Benson KF. Antioxidant and anti-inflammatory properties of an aqueous cyanophyta extract derived from arthrospira platensis: Contribution to bioactivities by the non-phycocyanin aqueous fraction. J. Med. Food. 2015; 18(5): 535-541. doi:10.1089/jmf.2014.0083

38. Elya B, Puspitasari N, Sudarmin AC. Antioxidant activity and inhibition of lipoxygenase activity ethanol extract of endosperm Arenga pinnata (Wurmb) merr. Asian J Pharm Clin Res. 2017; 10(Special Issue October): 76-80. doi:10.22159/ajpcr.2017.v10s5.23102

39. Ravi M, Tentu S, Baskar G, et al. Molecular mechanism of anti-cancer activity of phycocyanin in triple-negative breast cancer cells. BMC Cancer. 2015; 15(1): 1-13. doi:10.1186/s12885-015-1784-x

40. Jiang L, Wang Y, Yin Q, et al. Phycocyanin: A potential drug for cancer treatment. J. Cancer. 2017; 8(17): 3416-3429. doi:10.7150/jca.21058

41. Chakraborty P, Kumar S, Dutta D, Gupta V. Role of Antioxidants in Common Health Diseases. Res. J. Pharm. Tech. 2009; 2(2): 238-244.

42. Menon R. Antioxidants and their therapeutic potential- A review. Res. J. Pharm. Technol. 2013; 6(12): 1426-1429.

43. Ashfaq MH, Siddique A, Shahid S. Antioxidant Activity of Cinnamon zeylanicum: (A Review). Asian J. Pharm. Res. 2021; 11(2): 106-116. doi:10.52711/2231-5691.2021.00021

Received on 06.08.2023 Modified on 21.10.2023

Research J. Pharm. and Tech 2024; 17(7):3119-3126.

DOI: 10.52711/0974-360X.2024.00488